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HaloTag® Vectors for E. coli and Cell-Free Protein Expression

The following vectors are used for inducible expression of HaloTag® fusion proteins in E. coli and cell-free systems using the T7 RNA polymerase promoter. Expression levels depend highly on the nature of the protein, but in general the N-terminal HaloTag® fusion protein (e.g., pFN18A/K, Cat.# G2751, G2681) can increase expression level, enhance refolding and boost solubility of the expressed protein. HaloTag® vectors are supplied in two formats: as multiple cloning site (MCS) vectors for traditional cloning and as Flexi® System vectors.

The following vectors are used for inducible expression of HaloTag® fusion proteins in E. coli and cell-free systems using the T7 RNA polymerase promoter. Expression levels depend highly on the nature of the protein, but in general the N-terminal HaloTag® fusion protein (e.g., pFN18A/K, Cat.# G2751, G2681) can increase expression level, enhance refolding and boost solubility of the expressed protein. HaloTag® vectors are supplied in two formats: as multiple cloning site (MCS) vectors for traditional cloning and as Flexi® System vectors.

The Flexi® Vector System is a simple, directional cloning method for protein-coding sequences. It is based on two rare-cutting restriction enzymes, SgfI and PmeI, and provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi® Vectors without the need to resequence. Direct transfers can only occur between two N-terminal tagged vectors or from an N-terminal to a C-terminal vector. The MCS vectors and several Flexi® system vectors contain a His6-HaloTag® dual tag. The dual tag enables protein purification with the reusable Ni-resin while retaining the HaloTag® covalent labeling properties.

Note: Flexi® Vectors contain the lethal barnase gene to reduce background colonies without inserts during the subcloning procedure. Using the Flexi® Vector Cloning System replaces the barnase gene with your insert. These vectors, as purchased, cannot be cultured in normal laboratory strains of E. coli without an insert.

Features - Benefits

Choice of Systems: Choose between traditional (MCS) and Flexi® cloning to get the benefits of HaloTag® technology.

Dual Tag: Couple the protein solubility and labeling benefits of HaloTag® technology with the reusability and the throughput of Ni-affinity technology.

Versatile Cloning: Choose from a variety of expression systems and fusion tag orientations and then transfer to others as required (for Flexi® system).

Use Restrictions

Patents - Disclaimers

G7971, G8031, G2751, G2681, G1891, G1841, G1681, G1691, G8261, G8331, G8321, G8381 BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the researcher is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the researcher with a full refund. Researchers may use this product for research use only, no commercial use is allowed. Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence of the HaloTag® gene. Researchers may however clone heterologous DNA sequences at either or both ends of said HaloTag® gene so as to create fused gene sequences provided that the coding sequence of the resulting HaloTag® gene has no more than four (4) deoxynucleotides missing at the affected terminus when compared to the intact HaloTag® gene sequence. In addition, researchers must do one of the following in conjunction with use of the product: (1) use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties, or (2) contact Promega to obtain a license if Promega HaloTag® ligands are not to be used. Researchers may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license. With respect to any uses outside this label license, including any diagnostic, therapeutic or prophylactic uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THE PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

C8441, C8451, G2751, G2681, G1891, G1841, G1681, G1691, G8261, G8331, G8321, G8381 For research use only. Persons wishing to use this product or its derivatives in other fields of use, including without limitation, commercial sale, diagnostics or therapeutics, should contact Promega Corporation for licensing information.

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